Bis quaternary oximes



United States Patent 3,093,542 BIS QUATERNARY OXIMES Brennie E. Hackley, Jr., and Edward J. Poziomek, Edgewood, and George M. Steinherg, Baltimore, Md., assignors to the United States of America as represented by the Secretary of the Army No Drawing. Original application Apr. 28, 1959, Ser. No. 809,578, now Patent No. 3,077,476, dated Mar. 12, 1963. Divided and this application Oct. 11, 1961, Ser. No. 151,127

6 Claims. (Cl. 167-65) (Granted under Title 35, US. Code (1952), see. 266) This invention described herein may be manufactured and used by or for the Government of the United States of America for governmental purposes without the payment to us of any royalty thereon.

This is a division of application Serial No. 809,578, liled April 28, 1959, now Patent No. 3,077,476, granted March 12, 1963.

This invention is directed to certain diquaternary pyridinium halide oximes which are useful as chemotherapeutic and prophylactic agents for mammals poisoned by anticholinesterases, particularly the nerve gas known as GB or sarin, i.e., isopropyl methylphosphonofluoridate.

The invention relates to 1,1-polymethylene bis (4-formylpyridinium) halide dioximes wherein the polymethylene group contains from 2 to 6 carbon atoms. These compounds have the structural formula O H=N O H 'dioximes of the formula CH l 1 H-N 2X- N CH: C a l 7 H3 7 V i CH=NOH CH=NOH (II) The invention also relates to l,l(2-butenylene) bis (4 formylpyridinium) halide dioximes of the formula The invention further relates to 1,1-polymetl1ylene his (S-formylpyridinium) halide dioximes of the formula CH=NOH CH=NOH (IVa) wherein R is a polymethylene group containing from 2 to 6 carbon atoms.

Another class of compounds included are the 1,1'-(pice phenylenedimethylene) bis (3-formylpyridinium) halide dioximes of the formula I CH=NOH O H=N O H (IVb) This invention further relates to unsymmetrical bisquaternary 4-tormylpyridiniu-m halide m-onoximes of the fornm a N+ o 1 H93 N\ 2X- ([l=NOH o=NoH NH: NH: (VI) This invention also relates to the compounds (02115) aN-(CH!) F'N 2X- CH=NOH This invention also relates to the compounds N n (O 93 N\ 2X OH=NOH (VIII) While all these compounds are useful in varying degree for the purposes set out above, the different groups exhibit quite striking difierences in effectiveness. In all the above formulas, X- is chloride, bromide or iodide, which appear to be equivalents as to physiological action, except tor their efiect on solubility.

Wilson et al., in Patent No. 2,816,113, granted December 10, 1957, discloses a group of compounds which are efiective as antidotes for treatment of mammals poisoned by compounds of high anticholinesterase activity, such as the nerve gases diisopropylphosphorofluoridate ('DFP), isopnopyl methylphosphonofluoridate (GB), and O-ethyl, N,N dimethylphosphoroamidocyanidate (GA), as well as other related organic phosphorus compounds, includingmany. insecticides. The compound of the Wilson et al. group which has received by tar the most attention is Z-formyl-l-methyl pyradinium iodide oxime, commonly known as Z-pyridine aldoxi-me methiodide or Z-PAM. This compound is outstanding in its ability to reactivate, in yitro, acetyl cholinesterase which has been inhibited by, for example, GB. Thus, even as compared to the very closely related 4-PAM, disclosed in Example 11 of the Wilson patent, 2-PAM shows much greater activity. With isopropyl methylphosphonylated acetylcholinesterase the rate constant at pH 7.4 and 25 C. in the presence of 10- M acetylcholine is 2 10 per mole per minute tfor 2-PAM and 1.4)(10 per mole per minute for 4-PAM. The Wilson patent shows a high rate of survival in mice which had been poisoned with paraoxon and then treated with 2-PAM.

Nerve gas poisoning has been treated symptomatically with drugs which are pharmacologically antagonistic to acetyl choline. Such a compound is atropine and it is at present the recommended remedy. Recently, 2-PAM has been reported to enhance considerably the activity of atropine in the chemotherapeusis of poisoning due to organophosphorus compounds.

The compounds of Formula I above in which R contains from 2 to 6 carbon atoms are appreciably more effective than 2-PAM as reactivators of GB-inhibited acetylcholinesterase and also in enhancing the activity of atropine in both therapy and prophylaxis. When X is bromide, the variation of the rate constant for the in vitro reactivation of GB inhibited eel acetylcholinesterase at pH 7.4 and 25 C. was found to be as follows:

TABLE 1 R Rate constant (L/moles/minutes) M 7X10 (0112):. 6X10 (CH2)4 6x10 (CH2)5 1X1O4 (CH2)0 6X10 When administered in combination with atropine to animals poisoned with GB the order of effectiveness was somewhat different. Under these conditions the compound in which R: (CH-9 i.e., 1,1' trimethylene bis (4-formylphyridinium) bromide dioxime also known as TMB-4, was most effective. In rats challenged with a 2 LD dose of GB administered intravenously, all of a group of six animals survived if the atropine-TM-B-4 combination was administered intravenously immediately after poisoning. The atropine-Z-PAM combination saved only two of the group of animals. One the other hand, with dogs which were given a 20 LD dose of GB subcutaneously the survival ratios were the same (4/5) for the two treatments, which were given intravenously when symptoms appeared. However, the recovery time was much shorter for the surviving animals which received the TMB-4, i.e., 2 hours, as against 24 hours for those receiving the 2-PAM.

A summary of the reactivation rates and survival ratios for these compounds when administered therapeutically to rats together with atropine is as follows:

TABLE 2 R Reactivation Survival rate constant ratio (GB) 7x10 e/s e 10 6/6 6x10 6/6 1x10 6/6 (011m 6x10 3 4 These compounds constitute our presently preferred group.

Our compounds may be'employed prophylactically, i.e., injected before exposure to the anticholinesterase agent, e.g., GB, or therapeutically, i.e., injected subsequent to exposure.

The following series of experiments compares the eifectiveness of our presently preferred compound, TMB-4, with Z-PAM applied to various animals by these two To minimize absorption effects both the GB and TM'B-4 were ordinarily given intravenously. However, in the therapeutic tests on rabbits, dogs and cats, the GB was administered subcutaneously, since death from 20 LD intravenous dose of GB occurs so quickly that it is virtually impossible to give timely administration of the antidote.

Atropine, when administered, was included in the following amounts.

Mg./kg. Rats 4 Rabbits 2 Dogs and cats 0.5

The prophylactic doses were given within two minutes prior to the injection of the GB, the therapeutic" doses as soon as poisoning symptoms were visible.

Table 3 shows the results.

TABLE 3 A. PROPHYLAOTIC 2-PAR TMB-4 Survival ratio Survival ratio Amalsni Animals Dose mgjkg.

Dose WithmgJkg.

out atropine With atropine Without atropine With atropine Mice 40 Dogs Dogs The recovery periods, i.e., time for disappearance of symptoms of poisoning, among survivors in the above tests, with atropine, were as follows.

TABLE 4 2-PAM TM 13-4 Animals Prophylactic Therapeutic Prophylactic Therapeutic Rats Rabbits Cats Done The compounds of Formula I in which R contains from 7 to 10 carbon atoms are less effective than those of our preferred group. For these compounds the reactivation rate constant and the survival ratio for rats (measured as given above) were as follows, X- being bromide.

TABLE 5 Rate constant Survival ratio (GB) (CHaho While these compounds were ineffective in vivo against GB, they were, together with 2-PAM, very effective against certain other anticholinestenases, particularly that designated as VX by the US. Army Chemical Corps. All these compounds caused survival of all animals (survival rates of 4/4 and 6/6), when administered therapeutically to rats challenged by 2 LD doses of VX.

The compounds of Formula II exhibited properties intermediate those of the two subgroups of Formula I. When X- was chloride the compound had the following properties. (In this and tall following tables the survival ratios are those for rats challenged by 2 LD doses of GB or (VX) and the oxime was employed therapeuticall y.)

"Compounds of Formula 111 showed reactivation rates very close to those of our preferred group. Thus when X- in Formula III is bromide the reactivation rate constant was 8X10 as compared to the value for the R=(CH member of our preferred group of 6x10 For the unsaturated member (Formula III) the survival ratio for rats challenged by GB was only 1/4 as compared to 6/6 for the saturated analogue (Formula I). Both gave complete survival (ratios of 4/4 and 6/ 6) for animals challenged by VX, however.

Compounds of Formula IVa showed anomalous properties.

They gave reactivation rates which were low, but survival ratios which were high as compared to 2-PAM, as shown by Table 6, X- being bromide.

TABLE 6 R Reactivation Survival rate constant ratio (GB) (CHM 3. X10 4/4 (0119 12x10 3/4 The compounds of group IVb, which are closely related to those of IVa, were somewhat less eflective. When X- was bromide the compound had the following properties: Reactivation rate constant 2x10 survival ratio (GB)-2/ 4.

The compounds of group V were another group in which the results of therapeutic treatment against GB were better as compared to 2-PAM than the reactivation rate constants would suggest, as shown by the following table, X- being bromide.

The compounds of :Formulas VI, VII and VIII, while being of different structure are alike in exhibiting reactivation rate constants which are very low as compared to Z-PAM but giving high survival ratios as shown by Table 8, X- being bromide in each case.

TABLE 8 Formula No. Reactivation Survival 1 rate ratio (GB) VI 69 4/4 VII Negligible 4/4 VIII 67 4/4 Preparation of Compounds 4-pyridinecarboxaldehyde ox'ime was prepared by warming on a steam bath a neutralized aqueous solution of 4-pyridinecarboxaldehyde and hydroxylamine hydrochloride. The oxime had a melting point of 130-130.5 C. The 2- and 3-oximes were produced by similar methods.

The qua-ternization to produce dioximes was carried out by reacting the proper oxime with a 1, n dihaloal-hane (CH X employing a 3:1 molar ratio of oxime to halide. The unsymmetrical quaternary monoximes were obtained by reacting the pyridine oxime with the appropriate omega-halopropyl quaternary salt in a 1.5:1 molar ratio. 'Fwo procedures were utilized.

Procedure A: A mixture of the pyridine oxime and halide was dissolved in suflicient ethanol and refluxed for the period of time specified in Table9.

Procedure B: A mixture of the oxime and halide was dissolved in about 100 ml. of ethanol and heated in a 200 ml. capped pressure bottle (carbonated beverage type) for the length of time specified. The reaction mixtures were cooled to room temperature and the product removed by filtration. In several instances it was necessary to add absolute ether to effect complete precipitation. The products were recrystallized from ether. This procedure was usually employed because of its simplicity.

Table 9 gives the procedure, yields and melting points for representative compounds.

TABLE 9 For- Substituents Melting (m) mula Condi- Yield or decom- No. ticns percent position (d) Halide R R point, C.

I Br 35. 0 300 111. I Br 88. 2 238-241 (1. I B1 81. 0 239-241 (1. I B! 95.0 208-210 (1. I Br 8 h 85.0 219-223 (1.

II CI B, 68 hr 70 300 m. Iva... Br (CH2); 60 r 68 208-211 In. IVa Br (@119 60 hr 226-231 m. 1%.. B1 20 hr 83.5 248-251 in. V Br (CzH5)a B, 69 111:- 43 230-231 (1. V Br pyridine 64 hr 10 223-226 (1.

ring. VIII--. Br B, hr 16 201-203 d.

Further details regarding the preparation and properties of certain of our compounds are given in the following publications by us and our associates:

Pyridine Aldoximes, by Edward J. Poziomek, Brennie E. Hackley, Jr., and George M. Steinberg, Journal of Organic Chemistry, vol. 23, pp. 714-717 (May 1958); and Chemotherapeutic Eifectiveness of Trimethylene Bis (4-Formyl Pyrid-inium Bromide) Dioxime in Anticholinesterase Poisoning, by Edmund Bay, S. Kropp and L. F. Yates, Proceedings of the Society for Experimental Biology and Medicine, vol. 98, pages 107-109 (May 1958). These articles are to be considered incorporated by reference in this specification.

While we have shown a number of specific examples of compounds and their use, it will be obvious that various changes can be made without departing from our invention, which is defined by the following claims.

We claim:

1. A method of therapeutically treating a mammal which has been poisoned by a compound having high anticholinesterase activity which comprises injecting a composition consisting of atropine and a 1,1 polymethylene bis(4-formyl pyridinium) oxide halide compound of the formula EN) BT13 2X- CH=NOH H=NOH wherein R is a polymethylene group containing from two to six carbon atoms and X'- is selected from the class consisting of chloride, bromide and iodide.

2. A method of therapeutic-ally treating a mammal which has been poisoned by a compound of high anticholinesterase activity, comprising injecting a composition 5 consisting of atropine and 1,1-trimethylene bis (4-formy1- pyridinium) bromide dioxime.

7 3. A method of protecting a mammal against poisoning by a compound of high anticholinesterase activity comprising injecting, prior to said poisoning, Ia composition consisting of atropine and a 1,1 polymethylene bis(4-formyl pyridinium) oxime halide compound of the formula CH NOH OH=NOH wherein R is a polymethylene group containing from 2 to 6 carbon atoms and X- is selected from the class consisting of chloride, bromide and iodide.

4. A method of protecting a mammal against poisoning by a compound of high anticholinestepase activity, comprising injecting a composition consisting of atropine and 1,1-trimethylene bis-(4-formylpyridinium) bromide dioxime prior to said poisoning.

5. A method of thenapeutically treating a mammal which has been poisoned by a compound having high anticholinestemse activity which comprises injecting a composition consisting of atropine and a 1,1-po1ymethy1- ene bis(4-formyl pyridinium) oxime halide compound of 1' O N RN 2X- 0 O the formula ene bis-(3-formyl pyridinium) halide dioxime of the formula N R--N\ 2X CH=NOH CH=NOH wherein R is a polymethylene group containing from 2 to 6 carbon atoms and X is selected from the class consisting of chloride, bromide and iodide.

References Cited in the file of this patent Poziomek: Am. Chem. Soc. Abst. of Papers, 132nd Meeting, 1957, pages 16-0. 

1. A METHOD OF THERAPEUTICALLY TREATING A MAMMAL WHICH HAS BEEN POISONED BY A COMPOUND HAVING HIGH ANTICHOLINESTERASE ACTIVITY WHICH COMPRISES INJECTING A COMPOSITION CONSISTING OF ATROPINE AND A 1,1''-POLYMETHYLENE BIS(4-FORMYL PYRIDINIUM) OXIDE HALIDE COMPOUND OF THE FORMULA 